辐射介导下HeLa细胞中Cdc25B与IER5蛋白表达的关系

doi:10.3969/j.issn.1004-616x.2017.06.001

癌变·畸变·突变 ›› 2017, Vol. 29 ›› Issue (6): 405-410,417.doi: 10.3969/j.issn.1004-616x.2017.06.001

• 论著 • 下一篇

辐射介导下HeLa细胞中Cdc25B与IER5蛋白表达的关系

赵现哲¹, 刘晓丹², 周平坤², 姜晓燕¹, 丁库克¹

1. 中国疾病预防控制中心辐射防护与核安全医学所, 北京 100088;
2. 军事医学科学院放射与辐射医学研究所, 北京 100850

收稿日期:2017-02-21 修回日期:2017-06-02 出版日期:2017-11-30 发布日期:2017-11-30
通讯作者: 丁库克,E-mail:shouding@ccmu.edu.cn E-mail:shouding@ccmu.edu.cn
作者简介:赵现哲,E-mail:1241069549@qq.com。
基金资助:
国家自然科学基金（31170806，31640022，30770533，31770907）；北京市自然科学基金（7092013，7172146）

Expression of Cdc25B and IER5 in irradiated HeLa cells

ZHAO Xianzhe¹, LIU Xiaodan², ZHOU Pingkun², JIANG Xiaoyan¹, DING Kuke¹

1. National Institute for Radiological Protection, China CDC, Beijing 100088;
2. Institute of Radiation Medicine, Military Medical Science Academy of the PLA, Beijing 100850, China

Received:2017-02-21 Revised:2017-06-02 Online:2017-11-30 Published:2017-11-30

摘要/Abstract

摘要： 目的：探索辐射介导下HeLa细胞中Cdc25B与IER5蛋白表达的关系。方法：用4 Gy γ射线分别照射正常HeLa细胞和IER5-siRNA-HeLa细胞，收集照射后0、0.5、2、4、8、12、16、24、36 h的细胞样本，并设未照射细胞为对照组。采用qPCR法检测照射后Cdc25B mRNA的表达；Western blot检测细胞内IER5和Cdc25B蛋白的表达；流式细胞术检测照射后G2期细胞周期阻滞情况。结果：qPCR结果显示，照射后0.5~24 h内，与对照组相比，正常HeLa细胞内Cdc25B mRNA在0.5 h先下降2 h再上升之后再下降，差异均有统计学意义（P均 < 0.05）；在IER5-siRNA-HeLa细胞内Cdc25B mRNA在0.5 h先下降后再上升，差异均有统计学意义（P均 < 0.05，2 h组除外）。Western blot结果显示，照射后0.5~24 h内，与对照组相比，正常HeLa细胞内Cdc25B蛋白从2 h开始上升8 h开始下降，差异有统计学意义（P均 < 0.01）；IER5-siRNA-HeLa细胞内Cdc25B蛋白从0.5 h开始下降8 h开始逐渐上升，差异有统计学意义（P均 < 0.01，24 h除外）。照射后0~36 h内，与对照组相比，正常HeLa细胞与IER5-siRNA-HeLa细胞的IER5蛋白表达水平分别在照后8 h与0 h后开始出现明显上升，差异均具有统计学意义（P均 < 0.05）；而Cdc25B蛋白表达水分别在照射后8 h与0 h后开始出现下降，差异均具有统计学意义（P均 < 0.05），正常HeLa细胞与IER5-siRNA-HeLa细胞中Cdc25B蛋白与IER5蛋白的表达分别从8 h与0 h后开始呈现负相关关系（r分别为-0.740和-0.669，P均 < 0.01），当两种细胞中Cdc25B蛋白表达量下降时，G2期细胞周期阻滞比例升高（P < 0.05）。结论：辐射介导下HeLa细胞中Cdc25B蛋白表达变化是由Cdc25B mRNA变化引起的，并且Cdc25B与IER5蛋白表达呈负相关关系。

关键词: 辐射, IER5基因, Cdc25B, 细胞周期阻滞

Abstract: OBJECTIVE:The purpose of this study was to investigate the relationship between the expression of Cdc25B and IER5 proteins in irradiated HeLa cells. METHODS:HeLa cells and IER5-knockdown Hela cells were irradiated with 4 Gy of γ rays and were harvested at 0,0.5,2,4,8,12,16,24,and 36 h after irradiation. Expression of IER5 and Cdc25B proteins was examined by Western blot. G2 cell cycle arrest was examined by flow cytometry. Expression level of Cdc25B mRNA was examined by real-time quantitative PCR. Unexposed cells were used as controls. RESULTS:The qPCR results show that within 0.5-24 h after irradiation of the normal HeLa cells the Cdc25B mRNA was lower from 0.5 h the time-pointand then increased after 2 h,the difference was statistically significant (P < 0.05);In IER5 knockdown Hela cells,Cdc25B mRNA decreased first and then increased after 0.5 h,the difference was statistically significant (all P < 0.05,except 2 h). The Western blot results show that within 0.5-24 h after irradiation of normal HeLa cells,the Cdc25B protein began to rise from the 2 h time-point,then began to decrease at 8 h,the difference was statistically significant (all P < 0.05,except 0.5 h). In IER5 knockdown Hela cells,Cdc25B began to decrease from the 0.5 h time-point,then began to rise at 8 h gradually,the difference was statistically significant (all P < 0.05,except 24 h). Compared with the control group, IER5 protein was upregulated in two cells types after their irradiation and during the timepoints mentioned above. After radiation,the level of IER5 in HeLa began to increase significantly from the 8 h time-point while IER5 knockdown Hela cells from the 0 h. The differences were statistically significant (all P < 0.05). The level of Cdc25B in HeLa began to decline significantly from 8 h,while IER5 knockdown Hela cells from 0 h. The differences were statistically significant (P < 0.05). The protein expressions of Cdc25B and IER5 were negatively correlated at the 8 h and 0 h ime-points of radiation (R₁=-0.740,R₂=-0.669,P < 0.01,respectively),while the expression of Cdc25B was decreased. The ratio of G2 cell cycle arrest was increased in the two cells lines (P < 0.05). CONCLUSION:Expression of Cdc25B protein in HeLa cells was induced by Cdc25B mRNA changes,and the expression of Cdc25B was negatively related with the expression of IER5 protein.

Key words: radiation, IER5 gene, Cdc25B, cycle arrest

中图分类号:

R811.5

赵现哲, 刘晓丹, 周平坤, 姜晓燕, 丁库克. 辐射介导下HeLa细胞中Cdc25B与IER5蛋白表达的关系[J]. 癌变·畸变·突变, 2017, 29(6): 405-410,417.

ZHAO Xianzhe, LIU Xiaodan, ZHOU Pingkun, JIANG Xiaoyan, DING Kuke. Expression of Cdc25B and IER5 in irradiated HeLa cells[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2017, 29(6): 405-410,417.

参考文献

[1] MAX W,LYU M,YANG Y L,et al. IER5,a novel member of the slow-kinetics immediate-early genes[J]. Genomics,1999,55(3):327-334.
[2] WU X,PANG S T,SAHLIN L,et al. Gene expression profiling of the effects of castration and estrogen treatment in the rat uterus[J]. Biol Reprod,2003,69(4):1308-1317.
[3] SHI H M,DING K K,ZHOU P K,et al. Radiation-induced expression of IER5 is dose-dependent and not associated with the clinical outcomes of radiotherapy in cervical cancer[J]. Oncol Lett,2016,11:1309-1314.
[4] LI M J,WANG W W,CHEN S W,et al. Radiation dose effect of DNA repair-related gene expression in mouse white blood cells[J]. Med Sci Monit NLM,2011,17(10):290-297.
[5] PUJI A,TANYA P,CHARLOTTE W,et al. MAPK pathway activation delays G2/M progression by destabilizing Cdc25B[J]. J Biol Chem,2009,284(49):33781-33788.
[6] DING K K,SHANG H Z,HAO C,et al. Induced expression of the IER5 gene by γ ray irradiation and its involvement in cell cycle checkpoint control and survival[J]. Radiat Environ Bioph,2009,48(2):205-213.
[7] NAKAMURA S,NAGATA Y,TAN L,et al. Transcriptional repression of Cdc25B by IER5 inhibits the proliferation of leukemic progenitor cells through NF-YB and p300 in acute myeloid leukemia[J]. PLoS One,2011,116(11):1-15.
[8] 杨明月. 辐射后HepG2细胞IER5与CDC25B启动子相互作用[D]. 石家庄:河北医科大学,2014:27.
[9] DALVAI M,MONDESERT O,BOURDON J C,et al. Cdc25B is negatively regulated by p53 through Sp1 and NF-Y transcription factors[J]. Oncogene,2011,30:2282-2288.
[10] YOSHINORI A,TATSUYA K,ATSUSHI O,et al. IER5 generates anovel hypophosphorylated active from of HSF1 and contributes to tumorigenesis[J]. Scientific Rep RTS,2016,12:1-19.
[11] 吴玉梅,吉晨,丁库克,等. 编码IER5的SiRNA的DNA序列及其载体和应用:104726459A[P]. 2015-06-24.
[12] 丁库克,杨川杰,杨芬,等. 一种含有IER5基因的表达载体、单克隆细胞系及其应用:101525626A[P]. 2009-09-09.
[13] YANG J C,WANG Y L,DING K K,et al. IER5 promotes irradiation-and cisplatin-induced apoptosis in human hepatocellular carcinoma cells[J]. Am J Transl Res,2016,8(4):1789-1798.
[14] MATTHIEU L,BERNARD D,ANGEL R,et al. UV-induced downregulation of the CDC25B protein in human cells[J]. FEBS LETT,2010,584(6):1199-1204.
[15] BEATRIX B,ESTELLE S,BEMADETTE A,et al. Unschduled expression of Cdc25B in S-phsae leads to replicative stress and DNA damage[J]. Mol Cancer,2010,9:29.
[16] ROSE B,ODILE M,CORINNE L,et al. Cdc25B overexpression stabilises centrin 2 and promotes the formation of excess centriolar foci[J]. PLoS One,2013,8(7):e67822.
[17] WANG Z H,CLAES G T,VIVI A F,et al. Overexpression of Cdc25B,Cdc25C and phospho-Cdc25C(ser216) in vulvar squamous cell carcinomas are associated with malignant features and aggressive cancer phenotypes[J]. BMC Cancer,2010,10(233):1-11.
[18] ISLKAWA Y,SAKURAI H,KAWABATA S,et al. Heat-induced expression of the immediate-early gene IER5,and its involvement in the proliferation of heat-shocked cells[J]. FEBS J,2015,282(2):332-340.
[19] ISLKAWA Y,SAKURAI H,KAWABATA S,et al. HSF1 transcriptional activity is modulated by IER5 and PP2A/B55[J]. FEBS Lett,2015,589(10):1150-1155.
[20] KAWABATA S,ISHATA Y,ISLKAWA Y,et al. Immediate-early response 5(IER5) interacts with protein phosphatase 2A and regulates the phosphorylation of ribosomal protein S6 kinase and heat shock factor 1[J]. FEBS Lett,2015,589(23):3679-3685.
[21] PAN C Y,ZHU D H,WANG Y,et al. Human cytomegalovirus miR-UL 148D facilitates latent viral infection by targeting host cell immediate early response gene5[J]. PLoS One,2016,8:1-26.

辐射介导下HeLa细胞中Cdc25B与IER5蛋白表达的关系

Expression of Cdc25B and IER5 in irradiated HeLa cells

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